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Sample GSM591726 Query DataSets for GSM591726
Status Public on Dec 31, 2011
Title Day12 b
Sample type RNA
 
Source name cardiomyocyte-specific differentiated murine ES cell
Organism Mus musculus
Characteristics cell line: ES-D3 murine pluripotent cells
atcc catalog#: CRL-1934
time: day 12
Biomaterial provider Axiogenesis AG Germany
Treatment protocol ES cells were trypsinized (d0) and cultured in suspension to form embryoid bodies (EBs). Briefly, ES cells were transferred into Iscove's modified Dulbecco's Medium (IMDM) with 20% batch-tested FCS, nonessential amino acids (0.1 mM) and -mercaptoethanol (0.1 mM) (Invitrogen, Karlsruhe, Germany) in a 10 cm bacterial Petri dish and cultured on a shaker for 48 h. At d2, EBs were transferred into spinner flasks (Integra Cell Spin, IBS, Fernwald, Germany) and cultured for additional 7 days at 37°C, 5% CO2, and 95% humidity. Medium was exchanged at d5, d7, and d9; and at day 9 Puromyicn was added to select for cardiomyocytes. At d12, remaining cardiobodies were trypsinized to obtain a single cell suspension of cardiomyocytes (Cor.At cardiomyocytes). Cor.At cells were seeded onto a Fibronectin coated 10cm dish as described above, and cultured for 6h before extracting RNA.
Growth protocol aPIG 44 cells were cultured on mouse embroynic fibroblasts in high-glucose Dulbecco’s modified Eagle’s medium (DMEM) supplemented with nonessential amino acids (0.1 mM), L-glutamine (2 mM), beta-mercaptoethanol (0.1 mM), LIF (ESGR) (500 U/ml), neomycin (6 µg/mL), and batch-tested fetal calf serum (FCS) (15% v/v) (all Invitrogen, Karlsruhe, Germany).
Extracted molecule total RNA
Extraction protocol Total RNAs were extracted by using PeqGold RNApure (Peqlab Biotechnology, Erlangen, Germany) according to the manufacturer's instructions.
Label biotin
Label protocol Total RNA containing low molecular weight RNA was labelled using the flashtag RNA labeling kit (Genisphere, Hatfield, PA, USA) according to the manufacturer’s instructions.
 
Hybridization protocol Sample was hybridized to a GeneChip® miRNA Array (Affymetrix, Santa Claraq, CA, USA) at 48°C and 60 rpm for 16 hours then washed and stained on Fluidics Station 450 (Fluidics script FS450_0003).
Scan protocol Mircroarray was finally scanned on a GeneChip® Scanner 3000 7G (Affymetrix).
Description mouse ES cells (D3, ATCC CRL 1934) transfected with the alpha-MHC-Pac-IRES-EGFP vector
Data processing Data was normalized with RMA (Bioconductor, affy package).
Normalization and probe set summarization was carried out on the mouse-specific probe sets only.
 
Submission date Sep 09, 2010
Last update date Dec 31, 2011
Contact name Lin Gan
Organization name University Hospital Aachen
Department IZKF
Lab Genomics facility
Street address Pauwelsstr. 30
City Aachen
ZIP/Postal code 52074
Country Germany
 
Platform ID GPL8786
Series (1)
GSE24066 miRNA profiling during cardiomyocyte-specific differentiation of murine embryonic stem cells based on two different miRNA array platforms

Data table header descriptions
ID_REF
VALUE RMA-normalized signal intensity

Data table
ID_REF VALUE
mmu-let-7g_st 6.206011
mmu-let-7i_st 7.077992
mmu-miR-1_st 10.05116
mmu-miR-15b_st 8.124699
mmu-miR-23b_st 10.82934
mmu-miR-27b_st 7.90232
mmu-miR-29b_st 3.99708
mmu-miR-30a_st 6.921471
mmu-miR-30b_st 6.845895
mmu-miR-99a_st 4.158827
mmu-miR-99b_st 10.7669
mmu-miR-101a_st 4.009537
mmu-miR-124_st 4.178155
mmu-miR-125a-5p_st 10.66209
mmu-miR-125a-3p_st 5.129046
mmu-miR-125b-5p_st 9.488932
mmu-miR-126-5p_st 4.114345
mmu-miR-126-3p_st 4.994212
mmu-miR-127_st 10.20807
mmu-miR-128_st 5.813484

Total number of rows: 609

Table truncated, full table size 15 Kbytes.




Supplementary file Size Download File type/resource
GSM591726.CEL.gz 141.4 Kb (ftp)(http) CEL
Processed data included within Sample table

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